The Effect of Millisecond Pulsed Electric Fields (msPEF) on Intracellular Drug Transport with Negatively Charged Large Nanocarriers Made of Solid Lipid Nanoparticles (SLN): In Vitro Study

Abstract

Drug delivery technology is still a dynamically developing field of medicine. The main direction in nanotechnology research (nanocarriers, nanovehicles, etc.) is efficient drug delivery to target cells with simultaneous drug reduction concentration. However, nanotechnology trends in reducing the carrier sizes to several nanometers limit the volume of the loaded substance and may pose a danger of uncontrolled access into the cells. On the other hand, nanoparticles larger than 200 nm in diameter have difficulties to undergo rapid diffusional transport through cell membranes. The main advantage of large nanoparticles is higher drug encapsulation efficiency and the ability to deliver a wider array of drugs. Our present study contributes a new approach with large Tween 80 solid lipid nanoparticles SLN (i.e., hydrodynamic GM-SLN—glycerol monostearate, GM, as the lipid and ATO5-SLNs—glyceryl palmitostearate, ATO5, as the lipid) with diameters DH of 379.4 nm and 547 nm, respectively. They are used as drug carriers alone and in combination with electroporation (EP) induced by millisecond pulsed electric fields. We evaluate if EP can support the transport of large nanocarriers into cells. The study was performed with two cell lines: human colon adenocarcinoma LoVo and hamster ovarian fibroblastoid CHO-K1 with coumarin 6 (C6) as a fluorescent marker for encapsulation. The biological safety of the potential treatment procedure was evaluated with cell viability after their exposure to nanoparticles and EP. The EP efficacy was evaluated by FACS method. The impact on intracellular structure organization of cytoskeleton was visualized by CLSM method with alpha-actin and beta-tubulin. The obtained results indicate low cytotoxicity of both carrier types, free and loaded with C6. The evaluation of cytoskeleton proteins indicated no intracellular structure damage. The intracellular uptake and accumulation show that SLNs do not support transport of C6 coumarin. Only application of electroporation improved the transport of encapsulated and free C6 into both treated cell lines.Electronic supplementary materialThe online version of this article (doi:10.1007/s00232-016-9906-1) contains supplementary material, which is available to authorized users.

Nanoemulsion Structural Design in Co-Encapsulation of Hybrid Multifunctional Agents: Influence of the Smart PLGA Polymers on the Nanosystem-Enhanced Delivery and Electro-Photodynamic Treatment

“…The following parameters of electroporation were applied: electrical field intensity E (appl) = 500 V/cm, 5 rectangular unipolar pulses of 1 ms duration. The EP conditions were established according to our previous study [14]. The EP experiments were performed using Gene Pulser Xcell™ Electroporation System 165-2660 (BioRad).…”

“…Photocytotoxicity of NCs in SKOV-3 and CHO-K1 cells was measured after standard photodynamic procedure and PDT combined with the EP protocol described above by cellular mitochondrial activity determined by the (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) colorimetric assay according to the manufacturer’s procedure and our previous experiments [12,13,14,15]. The cells subject to PDT were irradiated after 24 h for 10 min with light in the range of 630–680 nm; the final energy delivered to the cell monolayer was 10 J/cm 2 .…”

“…Moreover, electroporation can be used for supported delivery or fusion of nanosystems containing multifunctional cargo, which is not possible with chemical methods. Thus, the EP method was effective not only for drugs and photosensitizers in free form but also for encapsulated ones [14]. Our previous study shows that EP enhances delivery and photodynamic activity of encapsulated cyanine IR-780 combined with flavonoids [15].…”

Benefitial role of electrochemotherapy in locally advanced pancreatic cancer – radiological perspective

“…The most commonly used is millisecond EP with relatively low voltage (up to 500 V/cm) to introduce ge netic material, lipid nanoparticles, and some drugs into the cells [36,37]. The nanosecond pulsed electroporation field (nsPEF) results in permeabilization of cell mem branes, including intracellular membranes with the loss of mitochondrial and nuclear membranes potential.…”

Research and development of the high-frequency square-wave pulse electroporation system

“…For electroporation application an individual duration of square-wave pulses is reported in range from approximately picoseconds to milliseconds. The submicrosecond (from picosecond to nanosecond) pulsed electric fields (nsPEF) pulses are used to induce electroporation of an intracellular organelle while the integrity of plasma membrane in a biological cell remains intact (Semenov et al 2015;Semenov et al 2016;Mao et al 2018;Pirc et al 2019;Tang et al 2020), while longer pulse up to milliseconds are used to enhance the transfer of large charged molecules (proteins, DNA, RNA) into the cells (Kulbacka et al 2016;Cervia et al 2018;Kašėta et al 2020).…”